Tommaso Renieri

9 + 0 immotile spermatozoa in an infertile man.

“9 + 0”‐unbewegliche Spermatozoen bei einem unfruchtbaren Mann

Further observations on the morphogenesis of the round headed human spermatozoa.

Sperm and testicular biopsies of an infertile human patient have been investigated using histochemical and electron microscopical technique. Spermiogram revealed a head defect, characterized with lacking acrosome and round and immature nucleoplasm, occuring in practically all cells and coiled tails in about a half of spermatozoa. EM study of spermatids has shown an abortive development of acrosome, whose primordium failed to attach the nucleus and expand, instead regressed. The nucleus failed in shaping and retarded in maturing, a disturbance apparently associated with the aplasia of hypoplasia of caudal manchette. The role of zinc in nuclear differentiation and the shape of head in the movement pattern are discussed.

Notulae seminologicae. 2. The "short tail" and "stump" defect in human spermatozoa.

Summary. In this note several cases of stunted tails involving the total sperm population in sterile humans are described. Half of the cases are classified as ‘short tailed’ spermatozoa, the other half as ‘stump defect’ previously described in bulls. Both defects are referred in details at electron microscopical level.

In vitro toxic effects of metal compounds on kinetic traits and ultrastructure of rabbit spermatozoa.

Metal compounds have been associated with male reproductive toxicity in vivo. The aim of the present study was to investigate the in vitro effects of 20 metal compounds using rabbit ejaculated spermatozoa as a study model for spermiotoxicity. Five of the metals tested (arsenic, cadmium, chromium, mercury and vanadium) reduced sperm motility and curvilinear velocity. Ultrastructural analyses revealed three types of damage to sperm head membranes in relation to the metal used: acrosome breakage with formation of various sized microvesicles (arsenic, cadmium, mercury and platinum); a large round hole (arsenic, cadmium and chromium), and numerous folds in the acrosome membrane (vanadium). The vanadium compound, followed by chromium and mercury compounds, determined a higher number of damaged spermatozoa. In conclusion, all the studied metal compounds, at levels higher than 1microM, may reduce sperm kinetic characteristics and probably fertilizing capacity by triggering specific morphological damages to the head and/or by inhibiting motility.

In vitro effect of gold and silver nanoparticles on human spermatozoa

The cytotoxicity of Au/Ag nanoparticles (NPs) on human spermatozoa was investigated in vitro. Semen from donors were incubated (37 °C, 60′–120′) with 30, 60, 125, 250 and 500 μM Au/Ag‐NPs. Sperm motility was evaluated following WHO guidelines; sperm viability was assessed with eosin Y test. Au‐NPs were characterised and localised with field emission gun‐based scanning transmission electron microscope/energy dispersive spectroscopy and transmission electron microscopy. Both tested NPs exerted a significant dose‐dependent effect on motility and viability of human spermatozoa (P < 0.001). Ag‐NPs seem to show a slightly elevated toxicity although not significant (P > 0.05). Au‐NPs were localised in spermatozoa, whereas Ag‐NPs were undetectable. In conclusion, Au‐NPs and Ag‐NPs do not appear to be harmful for human spermatozoa up to high concentrations (250–500 μM) that are probably difficult to reach in vivo. It is mandatory to explore the genotoxic effect of NPs in germ cells.

Effects of gold and silver nanoparticles in cultured human osteoarthritic chondrocytes.

The aim of our study was to evaluate the effects of gold (Au) and silver (Ag) nanoparticles (NPs) at different concentrations on cultured human osteoarthritic chondrocytes. Cell viability and inducible nitric oxide synthase expression were evaluated by light microscopy. Using transmission electron microscopy (TEM) and field emission gun‐based scanning transmission electron microscopy/energy dispersive spectroscopy (FEG‐STEM/EDS) allowed us to localize NPs. Gene expression of matrix metalloproteinases 1, 3 and 13 and A disintegrin and metalloproteinase with thrombospondin motifs ‐4 and ‐5 were carried out by real‐time polymerase chain reaction. A cell viability test indicated a significant dose‐dependent cytotoxic effect of both NPs. At concentrations of 160 and 250 μM NP light microscopy showed chondrocytes with signs of apoptosis and an increased presence of inducible nitric oxide synthase. Au‐NPs were characterized by FEG‐STEM/EDS and TEM analysis localized NPs in cytoplasm and in endocytotic vesicles. On the contrary, the Ag‐NPs were undetectable by FEG‐STEM/EDS and TEM. Increased gene expression, particularly in matrix metalloproteinase‐3, was observed for both NPs (160 μM), but at a concentration of 250 μM the expression of the evaluated genes became lower. Our in vitro studies, although preliminary, suggest that engineered Au and Ag‐NPs appear to be harmful for human osteoarthritic chondrocytes in high concentrations (160–250 μM). Copyright

Sperm with fibrous sheath dysplasia and anomalies in head-neck junction: focus on centriole and centrin 1.

Spermatozoa with a rare combination of two monomorphic sperm defects, dysplasia of the fibrous sheath (DFS) and alterations in head–mid‐piece junction were analysed. The main focus was to explore the status of the centriole, a key organisation during fertilisation, using the centrin 1, a calcium‐binding protein linked to this structure. The sperm quality was examined by light, scanning and transmission electron microscopy (SEM, TEM); immunocytochemistry was performed for tubulin, A‐kinase anchor protein 4 (AKAP4) and centrin 1. Spermatozoa showed DFS defect associated with anomalies in head–tail attachment detected by SEM and TEM. Immunolocalisation of tubulin, AKAP4 and centrin 1 confirmed these alterations. Centrin 1 was visible in 67% of spermatozoa (in only 13% centrin localised in a normal position); in the majority of sperm centrin 1s location was altered, sometimes bent; often four spots, indicating the presence of two implantation fossae, were detected. At the centriolar level, immunoreactive fragments, frequently invading the entire short and thick tail, were observed. Centrin 1 is an essential component of the spermatozoa connecting piece and plays a role in centrosome dynamics during sperm morphogenesis and in zygotes and early embryos during spindle assembly. It is important to shed light on these rare conditions in order to better manage the patients during assisted reproductive technology.

Abnormal elongation of midpiece, absence of axoneme and outer dense fibers at principal piece level, supernumerary microtubules: a sperm defect of possible genetic origin?

OBJECTIVE To characterize a flagellar defect involving 95% of the sperm population from an infertile man. DESIGN Case report. SETTING Interdepartmental Centre for Research and Therapy of Male Infertility, Siena, Italy. PATIENT(S) A 42-year-old infertile man with severe asthenozoospermia. INTERVENTION(S) Family history, physical examination, hormonal analysis, microbial assays, semen analysis, transmission electron microscopy (TEM) and scanning electron microscopy (SEM), tubulin distribution investigated by immunocytochemistry, fluorescence in situ hybridization for chromosomes 18, X, and Y. MAIN OUTCOME MEASURE(S) Ultrastructural abnormalities of the flagellum detected by methods listed. RESULT(S) Ultrastructural analysis revealed, in 95% of sperm cells, the total absence of the axoneme and outer dense fibers at the principal piece level, whereas the midpiece appeared abnormally long. Tubulin localization showed a total disorganization of the axoneme with a network of microtubular structures emerging randomly at any level of the flagellum. Fluorescence in situ hybridization analysis was normal. CONCLUSION(S) We report a rare sperm tail defect, characterized by abnormal elongation of the midpiece and absence of the axoneme and the outer dense fibers at the principal piece level in 95% of flagella. This defect occurs in the vast majority of the sperm population from a sterile man, and therefore a genetic origin could be hypothesized.

Notulae seminologicae. 1. New combinations of Kartagener's syndrome

Summary. In this note new consequences of the Kartageners syndrome are described. In males the syndrome involved a diffused sperm immaturity; in one female severe skeletal defects were present probably resulting from bad organization of the renal apparatus.

A case of severe asthenozoospermia: a novel sperm tail defect of possible genetic origin identified by electron microscopy and immunocytochemistry

OBJECTIVE To characterize a novel flagellar defect involving 98% of sperm tails. DESIGN Case report. SETTING Interdepartmental Centre for Research and Therapy of Male Infertility, Siena, Italy. PATIENT(S) A 45-year-old infertile man with severe asthenozoospermia. INTERVENTION(S) Family history, physical examination, hormonal analysis, microbial assays, semen analysis, transmission and scanning electron microscopy, tubulin distribution investigated by immunocytochemistry, fluorescence in situ hybridization (FISH) for chromosomes 9, 16, 18, X, and Y. MAIN OUTCOME MEASURE(S) Flagellar abnormalities detected by microscopical methods. RESULT(S) An apparent heterogeneity was observed: extremely elongated tails prone to ruptures; coiled tails at different levels with a strongly rolled axoneme or with a curl in the final flagellar segment; and V-shaped, isolated, bent tails. Transmission electron microscopy revealed the presence of normal heads, disorganized flagellar structures, and dynein deficiency. The FISH analysis was normal. CONCLUSION(S) We report a new sperm defect, characterized by abnormal elongation of the tail, which was prone to ruptures at different levels, concomitant with coiled tails, which were impossible to measure in length. This defect remained constant in different examined ejaculates and applied to the entire sperm population of a sterile man, the son of first-degree cousins, indicating a potential genetic origin.